The aim of this project is to develop and optimize our novel particle-based microarray platform, ArrayableESP, for application to high multiplex (50- to 2000-plex) proteomic analysis of human serum samples. ESPs are optically-encoded microfabricated particles that can be manipulated using magnetic force. They are manufactured on commercially available photolithographic equipment using efficient and robust techniques borrowed from the semi-conductor industry. They have significant advantages in cost throughput, scalability, and flexibility over existing bead-based liquid array platforms. We will use aptamers, instead of antibodies, on our platform, which will eliminate the need for sample pre-labeling/pre-processing and allow for a parallel method of homogeneous detection following protein binding. The combination of the ArrayableESP platform with aptamers is expected to increase accuracy of analysis and result in a simplified workflow. Such attributes, in combination with the nearly unlimited multiplex potential offered by the ArrayableESP platform, will provide a multiplex product with significant advantages over currently available technologies for proteomic analysis. ESP technology has broad applicability and should be a valuable tool in the diagnosis, prognosis, and characterization of a variety of disease states, including cancer, heart disease, arthritis and inflammatory diseases, kidney disease, liver disease, allergic responses, and infectious diseases (e.g. HIV). PUBLIC HEALTH RELEVANCE: Efficient high multiplexed genomic analysis has been achieved by others. Analysis of the proteome is expected to be the next important component of biological analysis for improving human health. Current methods for multiplex proteomic analysis are either unable to perform greater than 30-plex analysis, or at high multiplex require processes that result in inaccurate analysis with inefficient workflows. We are developing a cost-effective technology for low to high multiplex proteomic analysis with significant advantages over current comparable technologies. This technology should be useful in the diagnosis and prognosis of a variety of disease states, including cancer, heart disease, arthritis and inflammatory diseases, kidney disease, and infectious diseases (e.g. HIV).